Central Research Laboratory, Jilin University Bethune Second Hospital, Changchun 130041, PR China.
Oncol Rep. 2012 May;27(5):1481-7. doi: 10.3892/or.2012.1694. Epub 2012 Feb 21.
Evodiamine, an alkaloid isolated from Evodia rutaecarpa, possesses potent anticancer activity. Although many reports have elucidated the cytotoxic effects of evodiamine in a variety of cancer cells, little is known about the mechanism of evodiamine-induced cytotoxic activity in gastric cancer cells. To date, no report has addressed the synchronized role of autophagy and apoptosis in evodiamine-induced cytotoxic activity. This study was conducted to investigate the synchronized role of autophagy and apoptosis in evodiamine-induced cytotoxic activity on SGC-7901 human gastric adenocarcinoma cells and further to elucidate the underlying molecular mechanisms. The MTT assay was used to examine the cytotoxicity of evodiamine against SGC-7901 gastric adenocarcinoma cells. The effects of evodiamine on the cell cycle and apoptosis were measured by flow cyto-metry and cellular morphology was observed under a phase contrast microscope. Acridine orange (AO) staining was used to detect autophagy. The expression levels of Bcl-2 and Bax were detected by Western blotting. The expression level of Beclin‑1 in SGC-7901 cells was monitored by reverse transcription-polymerase chain reaction (RT-PCR). Here, we found that evodiamine significantly inhibited the proliferation of SGC-7901 cells and induced G2/M phase cell cycle arrest. Furthermore, both autophagy and apoptosis were activated during the evodiamine-induced death of SGC-7901 cells. Evodiamine-induced autophagy is partially involved in the death of SGC-7901 cells which was confirmed by using the autophagy inhibitor 3-methyladenine (3-MA). Additionally, Beclin-1 is involved in evodiamine-induced autophagy and the pro-apoptotic mechanisms of evodiamine may be associated with down-regulation of Bcl-2 and up-regulation of Bax expression. The inhibitory effects on SGC-7901 cells were associated with apoptosis, autophagy and cell cycle arrest at the G2/M phase in a dose-dependent manner. These results suggest that evodiamine is an effective natural compound for the treatment of gastric cancer and may represent a candidate for in vivo studies of monotherapies or combined antitumor therapies.
吴茱萸碱是从吴茱萸中分离得到的一种生物碱，具有很强的抗癌活性。虽然已有很多报道阐明了吴茱萸碱在多种癌细胞中的细胞毒性作用，但对于其在胃癌细胞中诱导细胞毒性的机制知之甚少。迄今为止，尚无报道涉及自噬和细胞凋亡在吴茱萸碱诱导的细胞毒性中的同步作用。本研究旨在探讨自噬和细胞凋亡在吴茱萸碱诱导的 SGC-7901 人胃腺癌细胞毒性中的同步作用，并进一步阐明其潜在的分子机制。MTT 法检测吴茱萸碱对 SGC-7901 胃腺癌细胞的细胞毒性。流式细胞术检测吴茱萸碱对细胞周期和凋亡的影响，相差显微镜观察细胞形态。吖啶橙（AO）染色检测自噬。Western blot 检测 Bcl-2 和 Bax 的表达水平。逆转录-聚合酶链反应（RT-PCR）检测 SGC-7901 细胞中 Beclin-1 的表达水平。结果发现，吴茱萸碱显著抑制 SGC-7901 细胞的增殖，诱导 G2/M 期细胞周期阻滞。此外，在吴茱萸碱诱导 SGC-7901 细胞死亡过程中，自噬和凋亡均被激活。自噬抑制剂 3-甲基腺嘌呤（3-MA）证实，吴茱萸碱诱导的自噬部分参与 SGC-7901 细胞的死亡。此外，Beclin-1 参与吴茱萸碱诱导的自噬，吴茱萸碱的促凋亡机制可能与 Bcl-2 下调和 Bax 表达上调有关。吴茱萸碱对 SGC-7901 细胞的抑制作用呈剂量依赖性，与细胞凋亡、自噬和 G2/M 期细胞周期阻滞有关。这些结果表明，吴茱萸碱是一种有效的胃癌治疗天然化合物，可能成为体内单药或联合抗肿瘤治疗研究的候选药物。
